Heterochromatin characterization of four fish species of the family Loricariidae (Siluriformes)

The karyotypic structures and the composition and distribution of the heterochromatin in the karyotypes of four catfish species belonging to four Loricariidae subfamilies were analysed, namely: Neoplecostomus microps (Neoplecostominae) with 2n=54 chromosomes, Harttia loricariformis (Loricariinae) with 2n=56 chromosomes, Hypostomus affinis (Hypostominae) with 2n=66 chromosomes and Upsilodus sp. (Upsilodinae) with 2n=96 chromosomes. The amount and composition of heterochromatin was quite unequal among the studied species, being copious and mainly GC-rich in Upsilodus sp. and scarce and balanced in H. loricariformis. All of the H. affinis heterochromatin is GC-rich and related with nucleolar organizing regions. N. microps show low quantity of interstitial and GC-rich heterochromatin, one of them being related with NORs. Trends in the macrokaryotypic diversification as well as in the distribution pattern of the heterochromatin are discussed.     

Enzyme Sets of Glycolysis, Gluconeogenesis, and Oxidative Pentose Phosphate Pathway Are Not Complete in Nongreen Highly Purified Amyloplasts of Sycamore (Acer pseudoplatanus L.) Cell Suspension Cultures

Differential centrifugation and Percoll-gradient centrifugation of protoplast lysates of suspension-cultured cells of sycamore (Acer pseudoplatanus L.) yielded pure amyloplasts. Contamination of the final amyloplast preparation by foreign compartments was assessed by measuring marker enzyme activities. The activity of alkaline pyrophosphatase was taken as a 100% plastid marker; relative to this marker, mitochondria (cytochrome c oxidase) averaged 0.34%, microbodies (catalase) 0.61%, and cytosol (alcohol dehydrogenase) 0.09%. Enzymatic activities of the glycolytic, gluconeogenic, pentose phosphate and the starch degradation pathways were found to be present in these amyloplast extracts in appreciable amounts. But the pyrophosphate-dependent phosphofructokinase and phosphoglyceromutase were judged to be essentially absent from amyloplasts because the activities of these enzymes were not enriched above the level of contaminating enzymatic activities in the amyloplast fractions. Additionally, the in vitro activities of starch phosphorylase, ATP dependent phosphofructokinase, NAD dependent glyceraldehyde-3 phosphate dehydrogenase, and glucose-6 phosphate dehydrogenase did not seem to support carbon fluxes from starch to triose phosphates as calculated from the rate of starch disappearance during carbon starvation of the cells. These results provide additional, indirect evidence for the recently emerged view that, in addition to the well known phosphate-triosephosphate translocator, another hexose phosphate and possibly also an ATP/ADP translocating system play major roles in nongreen plastids.     

Identifying critical migratory bottlenecks and high‐use areas for an endangered migratory soaring bird across three continents

Migrant birds face a number of threats throughout their annual cycle, including persecution, collision with energy infrastructure, and habitat and climate change. A key challenge for the conservation of migrants is the identification of important habitat, including migratory concentration areas, because species survival rates may be determined by events in geographically very limited areas. Remote‐tracking technology is facilitating the identification of such critical habitat, although the strategic identification of important sites and incorporation of such knowledge in conservation planning remains limited. We tracked 45 individuals of an endangered, soaring migrant (Egyptian vulture Neophron percnopterus), over 75 complete migrations that traversed three continents along the Red Sea Flyway. We summarize and contextualize migration statistics by season and age class, including migration start, midpoint, and end dates, as well as linear and cumulative migration distance, migration duration and speed, and route straightness. Then, using dynamic Brownian bridge movement models, we quantified space use to identify the most important migratory bottlenecks and high‐use areas on the flyway. These areas each accounted for < 5% of the overall movement range of the tracked birds, yet > 20% of all tracks passed through bottlenecks, and > 50% of the overall vulture time spent on migration fell within high‐use areas. The most important sites were located at the southeastern Red Sea coast and Bab‐el‐Mandeb Strait (Saudi Arabia, Yemen, Djibouti), the Suez Canal zone (Egypt), and the Gulf of Iskenderun (Turkey). Discouragingly however, none of the area within the major migratory bottlenecks was protected and < 13% of the high‐use areas were protected. This demonstrates a very concerning gap in the protected area network for migratory soaring birds along the Red Sea Flyway. Because reducing threats at migratory concentrations can be a very efficient approach to protect populations, our work provides clear guidelines where conservation investment is urgently needed to benefit as many as 35 migratory soaring‐bird species that regularly use the Red Sea Flyway.     

Stimulation of Symbiotic N2 Fixation in Trifolium repens L. under Elevated Atmospheric pCO2 in a Grassland Ecosystem

Symbiotic N2 fixation is one of the main processes that introduces N into terrestrial ecosystems. As such, it may be crucial for the sequestration of the extra C available in a world of continuously increasing atmospheric CO2 partial pressure (pCO2). The effect of elevated pCO2 (60 Pa) on symbiotic N2 fixation (15N-isotope dilution method) was investigated using Free-Air-CO2-Enrichment technology over a period of 3 years. Trifolium repens was cultivated either alone or together with Lolium perenne (a nonfixing reference crop) in mixed swards. Two different N fertilization levels and defoliation frequencies were applied. The total N yield increased consistently and the percentage of plant N derived from symbiotic N2 fixation increased significantly in T. repens under elevated pCO2. All additionally assimilated N was derived from symbiotic N2 fixation, not from the soil. In the mixtures exposed to elevated pCO2, an increased amount of symbiotically fixed N (+7.8, 8.2, and 6.2 g m-2 a-1 in 1993, 1994, and 1995, respectively) was introduced into the system. Increased N2 fixation is a competitive advantage for T. repens in mixed swards with pasture grasses and may be a crucial factor in maintaining the C:N ratio in the ecosystem as a whole.     

Does nitrogen nutrition restrict the CO2 response of fertile grassland lacking legumes?

The extent of the response of plant growth to atmospheric CO₂ enrichment depends on the availability of resources other than CO₂. An important growth-limiting resource under field conditions is nitrogen (N). N may, therefore, influence the CO₂ response of plants. The effect of elevated CO₂ (60 Pa) partial pressure (pCO₂) on the N nutrition of field-grown Lolium perenne swards, cultivated alone or in association with Trifolium repens, was investigated using free air carbon dioxide enrichment (FACE) technology over 3 years. The established grassland ecosystems were treated with two N fertilization levels and were defoliated at two frequencies. Under elevated pCO₂, the above-ground plant material of the L. perenne monoculture showed a consistent and significant decline in N concentration which, in general, led to a lower total annual N yield. Despite the decline in the critical N concentration (minimum N concentration required for non-N-limited biomass production) under elevated pCO₂, the index of N nutrition (ratio of actual N concentration and critical N concentration) was lower under elevated pCO₂ than under ambient pCO₂ in frequently defoliated L. perenne monocultures. Thus, we suggest that reduced N yield under elevated pCO₂ was evoked indirectly by a reduction of plant-available N. For L. perenne grown in association with T. repens and exposed to elevated pCO₂, there was an increase in the contribution of symbiotically fixed N to the total N yield of the grass. This can be explained by an increased apparent transfer of N from the associated N₂-fixing legume species to the non-fixing grass. The total annual N yield of the mixed grass/legume swards increased under elevated pCO₂. All the additional N yielded was due to symbiotically fixed N. Through the presence of an N₂-fixing plant species more symbiotically fixed N was introduced into the system and consequently helped to overcome N limitation under elevated pCO₂. Received: 11 November 1996 / Accepted: 20 May 1997     

Biogeographic patterns in the chromosomal distribution of a satellite DNA in the banded tetra Astyanax fasciatus (Teleostei: Characiformes)

The As-51 satellite DNA is a transposon-like sequence formerly described for arthropods and physically identifiable by fluorescent in situ hybridization. In the present work, we describe the occurrence of this sequence, as well the C-banding and karyotype composition, in populations of the group Astyanax fasciatus from Mogi-Guaçu (Araras-SP), Paranapanema (Angatuba and Pilar do Sul-SP), Ribeira de Iguape (Sete Barras-SP) and Tietê (Indaiatuba and Salesópolis-SP) river basins. The specimens from Sete Barras (10 M + 20SM + 12ST + 6A) and Araras (8 M + 22SM + 12ST + 6A) have 2n?=?48 chromosomes. The samples from Angatuba, Pilar do Sul and Indaiatuba presented 2n?=?46 chromosomes (12 M + 20SM + 10ST + 4A). The individuals collected in Salesópolis showed three cytotypes, bearing 2n?=?46 (12 M + 20SM + 10ST + 4A), 2n?=?48 (8 M + 22SM + 12ST + 6A) and 2n?=?50 (8 M + 16SM + 14ST + 12A). C-banding revealed large heterochromatic blocks at terminal chromosomal regions in all populations and/or cytotypes. All analyzed populations have conspicuous blocks carrying the As-51 satellite DNA, although the number of chromosomes bearing this repetitive sequence was variable among them. Such differences were not related to the diploid number of individuals, but rather to a biogeographic pattern. Aspects of the karyotype evolution and distribution of this sequence in distinct populations are discussed.     

Chromosome polymorphism in Astyanax fasciatus (Teleostei, Characidae), Part 3: Analysis of the RAPD and ISSR molecular markers

The Randomly Amplified Polymorphic DNA (RAPD) and Inter-Simple Sequence Repeats (ISSR) molecular markers were used to complement the study of chromosomal polymorphism in Astyanax fasciatus (Teleostei, Characidae) from the Mogi-Guaçu River (Southeastern Brazil), analyzed in three collection sites along the river (Ouro Fino – MG, Cachoeira de Emas – SP and Barrinha – SP). Two cytotypes (or karyotypic types), denominated standard cytotypes, were previously characterized, one including 2n = 46 chromosomes and the other 2n = 48 chromosomes, where all the chromosomes of the complement form homologous pairs. Additionally, variant karyotypic forms with 2n = 45, 46 and 47 chromosomes were also detected, although with a lower frequency in relation to the standard cytotypes. RAPD turned out little informative in the analysis of the observed situation, indicating a high value of migrants per generation among the cytotypes. On the other hand, ISSR showed a small structure, especially among the standard cytotypes from the Barrinha region where the Nm was 0.4301 with a genetic identity of 0.6862 and genetic distance of 0.3765. However, the general results obtained do not discard the possibility of interbreeding between both standard cytotypes and/or their descendants as a source of chromosome variation. The association between the cytogenetic and molecular markers viabilized putative explanatory scenery for the origin and evolution of the forms seen in A. fasciatus.     

Shade tolerance of <Emphasis Type="Italic">Ailanthus altissima</Emphasis> revisited: novel insights from southern Switzerland

The tree of heaven (Ailanthus altissima (Mill.) Swingle) is considered to be an early-successional, gap-obligate pioneer species with vigorous height growth, low shade tolerance, early fecundity and large seed production. It is a highly invasive species in many temperate and Mediterranean ecosystems outside its natural range, especially after disturbance. Due to its low shade tolerance, the potential of A. altissima to colonise undisturbed forests is thought to be low. In this study we analysed the potential of juvenile A. altissima to grow and survive in sweet chestnut (Castanea sativa Mill.) forests in southern Switzerland. We used hemispherical photography to assess the light conditions of 204 individuals of A. altissima (31 % generative, 69 % vegetative) aged between 1 and 7 years (median: 3 years) in six sites. Generative (seed-borne) and vegetative (clonal ramet) offspring of A. altissima are able to grow in light conditions well below the requirements of shade-intolerant tree species such as European larch (Larix decidua Mill.) and Scots pine (Pinus sylvestris L.). The relatively low light conditions found to be sufficient for the growth and survival of generative regeneration of A. altissima suggest a higher shade tolerance for this species than previously stated, at least for early regeneration. Consequently, the colonisation frontier of A. altissima should be intensively monitored in both forest openings but also in closed canopy forests in the vicinity of seed-bearing A. altissima.     

Acclimation of Photosynthesis to Elevated CO2 under Low-Nitrogen Nutrition Is Affected by the Capacity for Assimilate Utilization. Perennial Ryegrass under Free-Air CO2 Enrichment

Acclimation of photosynthesis to elevated CO₂ has previously been shown to be more pronounced when N supply is poor. Is this a direct effect of N or an indirect effect of N by limiting the development of sinks for photoassimilate? This question was tested by growing a perennial ryegrass (Lolium perenne) in the field under elevated (60 Pa) and current (36 Pa) partial pressures of CO₂ (pCO2) at low and high levels of N fertilization. Cutting of this herbage crop at 4- to 8-week intervals removed about 80% of the canopy, therefore decreasing the ratio of photosynthetic area to sinks for photoassimilate. Leaf photosynthesis, in vivo carboxylation capacity, carbohydrate, N, ribulose-1,5-bisphosphate carboxylase/oxygenase, sedoheptulose-1,7-bisphosphatase, and chloroplastic fructose-1,6-bisphosphatase levels were determined for mature lamina during two consecutive summers. Just before the cut, when the canopy was relatively large, growth at elevated pCO₂ and low N resulted in significant decreases in carboxylation capacity and the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase protein. In high N there were no significant decreases in carboxylation capacity or proteins, but chloroplastic fructose-1,6-bisphosphatase protein levels increased significantly. Elevated pCO₂ resulted in a marked and significant increase in leaf carbohydrate content at low N, but had no effect at high N. This acclimation at low N was absent after the harvest, when the canopy size was small. These results suggest that acclimation under low N is caused by limitation of sink development rather than being a direct effect of N supply on photosynthesis.     

Evidence that P deficiency induces N feedback regulation of symbiotic N2 fixation in white clover (Trifolium repens L.)

Trifolium repens L. was grown to test the following hypotheses: when P is deficient (i) N2 fixation decreases as a result of the plant's adaptation to the low N demand, regulated by an N feedback mechanism, and (ii) the decrease in the photosynthetic capacity of the leaves does not limit N2 fixation. Severe P deficiency prevented nodulation or stopped nodule growth when the P deficiency occurred after the plants had formed nodules. At low P, the proportion of whole-plant-N derived from symbiotic N2 fixation decreased, whereas specific N2 fixation increased and compensated partially for poor nodulation. Leaf photosynthesis was reduced under P deficiency due to low Vc,max and Jmax. Poor growth or poor performance of the nodules was not due to C limitation, because (i) the improved photosynthetic performance at elevated pCO2 had no effect on the growth and functioning of the nodules, (ii) starch accumulated in the leaves, particularly under elevated pCO2, and (iii) the concentration of WSC in the nodules was highest under P deficiency. Under severe P deficiency, the concentrations of whole-plant-N and leaf-N were the highest, indicating that the assimilation of N exceeded the amount of N required by the plant for growth. This was clearly demonstrated by a strong increase in asparagine concentrations in the roots and nodules under low P supply. This indicates that nodulation and the proportion of N derived from symbiotic N2 fixation are down-regulated by an N feedback mechanism.